. 2018 May 1;163(2):639–654. doi: 10.1093/toxsci/kfy066

Table 2.

The Percentage of Immune Cells in Immunologically Naïve Offspring

	Female 0.1 µg/ml versus Control			Male 0.1 µg/ml versus Control			F versus M	Female 1 µg/ml versus Control		Male 1 µg/ml versus Control		F versus M
	Control	0.1 µg/ml	p-Value^a	Control	0.1 µg/ml	p-Value^a	p-Value^b	1 µg/ml	p-Value^a	1 µg/ml	p-Value^a	p-Value^b
Bone marrow
HSC	0.0007 ± 0.0002	0.0011 ± 0.0002	.51	0.0015 ± 0.0004	0.0016 ± 0.0002	.96	.07	0.003 ± 0.001	.003*	0.0017 ± 0.0003	.91	.49
LSK	0.014 ± 0.001	0.014 ± 0.005	.99	0.014 ± 0.002	0.012 ± 0.001	.69	.98	0.015 ± 0.002	.96	0.015 ± 0.002	.92	.97
LK	0.30 ± 0.02	0.33 ± 0.06	.82	0.34± 0.02	0.35 ± 0.03	.95	.37	0.41 ± 0.01	.26	0.36 ± 0.03	.85	.53
MPP	0.0117 ± 0.0001	0.011 ± 0.004	.98	0.011 ± 0.001	0.008 ± 0.001	.42	.68	0.009 ± 0.002	.79	0.011 ± 0.002	.89	.61
Lineage ^-	2.7 ± 0.17	2.4 ± 0.64	.90	2.7 ± 0.28	2.5 ± 0.24	.84	.55	3.68 ± 0.13	.31	2.9 ± 0.12	.76	.15
CLP	0.04 ± 0.01	0.04 ± 0.02	.99	0.039 ± 0.004	0.03 ± 0.01	.59	.77	0.002 ± 0.001	.25	0.033 ± 0.003	.58	.01*
GMP	0.026 ± 0.004	0.044 ± 0.001	.03*	0.04 ± 0.01	0.04 ± 0.01	.96	1.0	0.05 ± 0.01	.04*	0.038 ± 0.002	.99	.30
Pre-GM	0.070 ± 0.003	0.08 ± 0.02	.88	0.09 ± 0.01	0.10 ± 0.01	.80	.09	0.066 ± 0.002	.95	0.11 ± 0.01	.69	.08
Pre-MegE	0.13 ± 0.01	0.13 ± 0.02	.99	0.120 ± 0.004	0.12 ± 0.01	.95	.55	0.1700 ± 0.00	.19	0.12± 0.01	.95	.03*
Thymus
DN1	0.76 ± 0.02	1.6 ± 0.86	.41	0.77 ± 0.09	0.76 ± 0.07	.98	.30	0.51 ± 0.06	.93	0.77 ± 0.08	.99	.07
DN2	0.12 ± 0.01	0.17 ± 0.02	.01*	0.09 ± 0.01	0.09 ± 0.01	.97	.003*	0.07 ± 0.01	.06	0.11 ± 0.01	.86	.47
DN3	1.25 ± 0.062	1.3 ± 0.09	.99	1.14 ± 0.039	1.15 ± 0.040	.99	.33	1.01 ± 0.005	.06	1.21 ± 0.033	.43	.18
DN4	0.72 ± 0.09	5.9 ± 4.9	.36	1.1 ± 0.21	1.04 ± 0.139	.95	.29	0.52 ± 0.07	.99	1.04 ± 0.144	.95	.06
DP	49.2 ± 3.67	37.6 ± 16.1	.62	50.5 ± 1.62	50.9 ± 4.31	.99	.44	39.5 ± 4.55	.76	53.7 ± 3.35	.73	.08
DP CD3⁺	0.03 ± 0.01	0.03 ± 0.01	.85	0.12 ± 0.04	0.08 ± 0.01	.53	.06	0.017 ± 0.004	.49	0.12 ± 0.02	.99	.01*
TCRβ⁺	5.3 ± 0.67	14.6 ± 6.4	.19	8.2 ± 0.98	7.1 ± 0.43	.54	.22	3.8 ± 0.19	.95	7.5 ± 0.86	.73	.04*
CD4⁺	4.2 ± 0.27	7.8 ± 2.4	.17	5.7 ± 0.48	5.6 ± 0.39	.94	.32	3.6 ± 0.34	.95	5.5 ± 0.51	.9	.07
CD8⁺	0.54 ± 0.09	0.26 ± 0.05	.36	1.1 ± 0.33	0.91 ± 0.18	.83	.30	0.26 ± 0.05	.99	0.93 ± 0.14	.86	.02*
TCRγδ⁺	0.36 ± 0.02	0.55 ± 0.17	.34	0.45 ± 0.05	0.45 ± 0.05	.99	.55	0.27 ± 0.03	.78	0.51 ± 0.08	.73	.04*
Treg	0.095 ± 0.004	0.63 ± 0.51	.37	0.13 ± 0.02	0.12 ± 0.02	.94	.29	0.08 ± 0.01	.99	0.11 ± 0.01	.66	.06
Spleen
CD19⁺	48.3 ± 3.16	49.1 ± 3.45	.98	47.2 ± 8.41	53.6 ± 1.19	.59	.22	54.5 ± 4.45	.47	51.5 ± 1.61	.77	.47
CD4⁺	10.8 ± 1.51	12.3 ± 0.984	.66	12.2 ± 1.69	13.7 ± 0.492	.51	.23	9.6 ± 0.39	.81	12.9 ± 0.17	.81	.001*
CD8⁺	7.8 ± 0.39	9.03 ± 0.087	.06	8.3 ± 0.47	9.2 ± 0.19	.09	.49	5.9 ± 0.16	.02*	8.4 ± 0.09	.92	.0001*
NK1.1⁺	2.2 ± 0.18	1.9 ± 0.24	.62	2.8 ± 0.44	2.1 ± 0.14	.16	.55	1.5 ± 0.06	.13	2.4 ± 0.09	.41	.004*
TCRγδ⁺	1.6 ± 0.07	1.5 ± 0.11	.72	1.5 ± 0.06	1.49 ± 0.025	.92	.62	1.36 ± 0.030	.16	1.4 ± 0.08	.49	.61
Treg	0.59 ± 0.14	0.71 ± 0.06	.69	0.78 ± 0.20	0.86 ± 0.07	.89	.21	0.61 ± 0.09	.99	0.85 ± 0.04	.91	.04*
F4/80⁺	3.3 ± 0.36	4.9 ± 1.9	.49	2.8 ± 0.23	3.1 ± 0.13	.29	.30	5.2 ± 0.32	.50	3.1 ± 0.11	.25	.002*
Gr-1⁺	5.8 ± 0.31	8.8 ± 2.2	.24	5.9 ± 0.61	6.6 ± 0.20	.33	.31	7.5 ± 0.67	.63	6.4 ± 0.07	.52	.06
Gr-1⁺CD11b⁺	0.53 ± 0.05	2.56 ± 2.01	.38	0.42 ± 0.02	0.44 ± 0.02	.82	.26	1.9 ± 0.19	.66	0.46 ± 0.03	.41	.0002*
CD11c⁺MHCII^hi	1.37 ± 0.044	1.38 ± 0.035	.98	1.2 ± 0.19	1.3 ± 0.07	.77	.42	1.2 ± 0.10	.23	1.3 ± 0.09	.86	.74
CD11b⁺DCs	0.15 ± 0.03	0.20 ± 0.05	.39	0.16 ± 0.02	0.17 ± 0.01	.63	.32	0.11 ± 0.01	.51	0.17 ± 0.01	.72	.03*
CD103⁺DCs	0.025 ± 0.004	0.04 ± 0.01	.24	0.03 ± 0.01	0.02 ± 0.01	.81	.22	0.02 ± 0.01	0.99	0.03 ± 0.01	.97	.51
Lymph nodes
NK1.1⁺	0.44 ± 0.05	0.34 ± 0.06	.38	0.41 ± 0.07	0.36 ± 0.05	.51	.75	0.45 ± 0.04	.99	0.29 ± 0.03	.86	.04*
CD19⁺	34.80 ± 6.010	28.6 ± 9.17	.77	27.4 ± 1.69	32.3 ± 5.24	.99	.73	43.4 ± 2.55	.68	29.6 ± 1.56	.89	.01*
TCRγδ⁺	1.8 ± 0.31	1.7 ± 0.29	.96	2.3 ± 0.70	1.1 ± 0.04	.90	.08	1.06 ± 0.005	.28	1.04 ± 0.068	.85	.87
CD4⁺	19.7 ± 2.56	16.3 ± 5.48	.76	16.5 ± 1.43	16.6 ± 3.62	.79	.97	23.9 ± 0.800	.70	15.2 ± 1.36	.27	.01*
CD8⁺	14.6 ± 1.51	13.2 ± 4.18	.91	12.4 ± 1.98	13.6 ± 3.16	.13	.94	16.3 ± 0.600	.88	10.9 ± 0.959	.11	.02*
Treg	1.5 ± 0.13	1.1 ± 0.38	.49	1.4 ± 0.37	1.5 ± 0.45	.99	.57	1.9 ± 0.12	.46	1.2 ± 0.18	.89	.07
F4/80⁺	0.72 ± 0.11	0.92 ± 0.23	.60	0.55 ± 0.07	0.55 ± 0.07	.86	.20	0.60 ± 0.03	.85	0.51 ± 0.06	.89	.36
Gr-1⁺	5.9 ± 0.68	5.9 ± 1.4	.99	5.4 ± 0.72	5.8 ± 1.1	.91	.95	9.5 ± 0.20	.09	4.7 ± 0.32	.8	.001*
Gr-1⁺CD11b⁺	0.07 ± 0.01	0.18 ± 0.13	.55	0.05 ± 0.01	0.05 ± 0.01	.99	.29	0.14 ± 0.05	.81	0.038 ± 0.003	.77	.03*
CD11c⁺MHCII^hi	0.85 ± 0.18	0.89 ± 0.07	.97	0.61 ± 0.06	0.71 ± 0.08	.41	.16	0.96 ± 0.08	.86	0.63 ± 0.04	.94	.02*
CD11b⁺DCs	0.23 ± 0.04	0.34 ± 0.14	.57	0.14 ± 0.03	0.18 ± 0.03	.53	.26	0.16 ± 0.02	.83	0.18 ± 0.03	.53	.62
CD103⁺DCs	0.19 ± 0.05	0.17 ± 0.07	.95	0.13 ± 0.02	0.14 ± 0.03	.95	.67	0.19 ± 0.01	.99	0.11 ± 0.01	.69	.01*

The mean percentage (±SEM) of the indicated cell types in primary and secondary immune organs from female and male offspring in each treatment group. Cells were collected from the bone marrow, thymus, and lymph nodes of immunologically naïve offspring. There were 4–8 offspring of each sex in each exposure group, and same-sex offspring within each exposure group were from different dams. Cell populations in the bone marrow include: HSC (Lineage[Lin]^negSca-1⁺cKit⁺CD105^hiCD150⁺); LSK cells (Lin^negSca-1⁺Kit⁺ cells); LK cells (Lin^negKit⁺ cells); multipotent progenitors (MPP; Lineage^negSca-1⁺cKit⁺CD105^intCD150^neg); common lymphocyte progenitors (CLP; Lineage^negSca-1⁺cKit⁺CD105^hiCD150⁺CD127⁺); granulocyte monocyte progenitors (GMP; Lin^negKit⁺CD16/32⁺CD150^neg); pre-granulocyte/monocyte cells (pre-GM; Lin^negKit⁺CD16/32^negCD150^negCD105^int); pre-megakaryocyte/erythrocyte cells (pre-MegE; Lin^negKit⁺CD16/32^negCD150⁺CD105^hi). In the thymus, cells were defined as double negative: DN1 (CD44⁺CD25^negCD4^negCD8^neg); DN2 (CD44⁺CD25⁺CD4^negCD8^neg); DN3 (CD44^negCD25⁺CD4^negCD8^neg); DN4 (CD44^negCD25^negCD4^negCD8^neg); double positive (DP; CD3⁺CD4⁺CD8⁺); CD8 SP (CD3⁺CD4^negCD8⁺); CD4 SP (CD3⁺CD4⁺CD8^neg); Tregs (Foxp3⁺CD25⁺CD4⁺CD8^negCD3⁺); γδT cells (γδTCR⁺CD3⁺CD4^-CD8^-). In the lymph node, cells were defined as: CD8⁺ T cells (CD3⁺CD8⁺); CD4⁺ T cells (CD3⁺CD4⁺); Tregs (Foxp3⁺CD25⁺CD4⁺); T cells (γδTCR⁺CD3⁺CD4^-CD8^-); neutrophils (Gr-1⁺); macrophages (F4/80⁺); dendritic cells (DCs; CD11c⁺MHCII^hi), CD11b⁺DCs (CD11b⁺CD103-CD11c⁺MHCII^hi cells), and CD103⁺DCs (CD11b^-CD103⁺CD11c⁺MHCII^hi cells); NK cells (CD3^negNK1.1⁺); B cells (CD3^negCD19⁺).

The p-value for mixture-exposed group compared with control group within the indicated sex.

The p-value for female versus male offspring from dams treated with the indicated concentration of the mixture.

Denotes p ≤.05.